A5062158055- Parkinson's Disease Mechanisms and Treatments
- Alzheimer's disease research and treatments
- Genetic Neurodegenerative Diseases
- Neurological disorders and treatments
- Muscle Physiology and Disorders
- Prion Diseases and Protein Misfolding
- Conducting polymers and applications
- Nuclear Receptors and Signaling
- Monoclonal and Polyclonal Antibodies Research
- Neurological diseases and metabolism
- Botulinum Toxin and Related Neurological Disorders
- Mitochondrial Function and Pathology
- Thyroid Disorders and Treatments
- Cholinesterase and Neurodegenerative Diseases
- Ion channel regulation and function
- Biotin and Related Studies
- Protein Structure and Dynamics
- Neurogenetic and Muscular Disorders Research
- Advanced Fluorescence Microscopy Techniques
- Multiple Myeloma Research and Treatments
- Nanofabrication and Lithography Techniques
- Flow Measurement and Analysis
- Cancer-related gene regulation
- Cell Image Analysis Techniques
- Oceanographic and Atmospheric Processes
Instituto di Biofisica
2012-2023
National Research Council
2010-2023
Dulbecco Telethon Institute
2017-2023
University of Trento
2017-2023
University of Cambridge
2012-2022
Human Technopole
2022
Fondazione Bruno Kessler
2012-2020
University of Padua
2006-2012
Indiana University – Purdue University Indianapolis
2006
Molecular Kinetics (United States)
2006
Significance Growing experimental evidence suggests that the pathological spreading of alpha-synuclein aggregates in Parkinson’s disease is mediated through a process templated seeding whereby catalyze conversion soluble protein molecules into their aggregated forms. A molecular-level understanding this still lacking. Here, we determine concentrations and numbers necessary for effective alpha-synuclein, thus providing quantitative framework to understand conditions when its seeded...
Abstract Super-resolution microscopy allows biological systems to be studied at the nanoscale, but has been restricted providing only positional information. Here, we show that it is possible perform multi-dimensional super-resolution imaging determine both position and environmental properties of single-molecule fluorescent emitters. The method presented here exploits solvatochromic fluorogenic nile red extract emission spectrum each dye molecule simultaneously enabling mapping...
Abstract Aggregation of alpha-synuclein (α-Syn) drives Parkinson’s disease (PD), although the initial stages self-assembly and structural conversion have not been directly observed inside neurons. In this study, we tracked intracellular conformational states α-Syn using a single-molecule Förster resonance energy transfer (smFRET) biosensor, show here that converts from monomeric state into two distinct oligomeric in neurons concentration-dependent sequence-specific manner. Three-dimensional...
The misfolding and aggregation of proteins into amyloid fibrils characterizes many neurodegenerative disorders such as Parkinson's Alzheimer's diseases. We report here a method, termed SAVE (single aggregate visualization by enhancement) imaging, for the ultrasensitive detection individual oligomers using single-molecule fluorescence microscopy. demonstrate that this method is able to detect presence aggregates α-synuclein, tau, amyloid-β. In addition, we show can also be identified in human...
Abstract Oligomers of alpha-synuclein are toxic to cells and have been proposed play a key role in the etiopathogenesis Parkinson’s disease. As certain missense mutations gene encoding for induce early-onset forms disease, it has suggested that these variants might an inherent tendency produce high concentrations oligomers during aggregation, although direct experimental evidence this is still missing. We used single-molecule Förster Resonance Energy Transfer visualize directly protein...
α-Synuclein oligomers can be toxic to cells and may responsible for cell death in Parkinson's disease. Their typically low abundance highly heterogeneous nature, however, make such species challenging study using traditional biochemical techniques. By combining fast-flow microfluidics with single-molecule fluorescence, we are able rapidly follow the process by which of αS formed characterize themselves. We have used technique show that populations different FRET efficiencies varying...
Familial and idiopathic Parkinson's disease (PD) is associated with the abnormal neuronal accumulation of α-synuclein (aS) leading to β-sheet-rich aggregates called Lewy Bodies (LBs). Moreover, single point mutation in aS gene multiplication lead autosomal dominant forms PD. A connection between PD 14-3-3 chaperone-like proteins was recently proposed, based on fact that some isoforms can interact genetic PD-associated such as parkin, LRRK2 were found components LBs human In particular, a...
The aberrant aggregation of α-synuclein is associated with several human diseases, collectively termed the α-synucleinopathies, which includes Parkinson's disease. progression these diseases is, in part, mediated by extracellular oligomers that may exert effects through mechanisms, including prion-like transfer, direct cytotoxicity, and pro-inflammatory actions. In this study, we show two abundant chaperones, clusterin α2-macroglobulin, directly bind to exposed hydrophobic regions on surface...
Abstract The protein alpha-synuclein (αS) self-assembles into toxic beta-sheet aggregates in Parkinson’s disease, while it is proposed that αS forms soluble alpha-helical multimers healthy neurons. Here, we have made vitro using arachidonic acid (ARA), one of the most abundant fatty acids brain, and characterized them by a combination bulk experiments single-molecule Fӧrster resonance energy transfer (sm-FRET) measurements. data suggest ARA-induced oligomers are alpha-helical, resistant to...
Abstract Recent computational advancements in the simulation of biochemical processes allow investigating mechanisms involved protein regulation with realistic physics-based models, at an atomistic level resolution. These techniques allowed us to design a drug discovery approach, named Pharmacological Protein Inactivation by Folding Intermediate Targeting (PPI-FIT), based on rationale negatively regulating levels targeting folding intermediates. Here, PPI-FIT was tested for first time...
Abstract Spinobulbar muscular atrophy (SBMA) is caused by CAG expansions in the androgen receptor gene. Androgen binding to polyQ-expanded triggers SBMA through a combination of toxic gain-of-function and loss-of-function mechanisms. Leveraging cell lines, mice, patient-derived specimens, we show that co-regulators lysine-specific demethylase 1 (LSD1) protein arginine methyltransferase 6 (PRMT6) are overexpressed an androgen-dependent manner specifically skeletal muscle patients mice. LSD1...
Spinal and bulbar muscular atrophy (SBMA) is a neuromuscular disorder caused by polyglutamine expansion in the androgen receptor (AR) characterized loss of lower motor neurons. Here we investigated pathological processes occurring muscle biopsy specimens derived from SBMA patients and, as controls, age-matched healthy subjects suffering amyotrophic lateral sclerosis (ALS) neurogenic atrophy. We detected atrophic fibers SBMA, ALS patients. In addition, was presence large number hypertrophic...
Abstract α‐synuclein (α‐Syn) is an abundant brain protein whose mutations have been linked to early‐onset Parkinson's disease (PD). We recently demonstrated, by means of a single‐molecule force spectroscopy (SMFS) methodology, that the conformational equilibrium monomeric wild‐type (WT) α‐Syn shifts toward β‐containing structures in several unrelated conditions PD pathogenicity. Herein, we follow same methodology previously employed for WT characterize heterogeneity pathological mutants...
Small oligomers of the protein α-synuclein (αS) are highly cytotoxic species associated with Parkinson's disease (PD). In addition, αS can form co-aggregates its mutational variants and other proteins such as amyloid-β (Aβ) tau, which implicated in Alzheimer's disease. The processes self-oligomerization co-oligomerization are, however, challenging to study quantitatively. Here, we have utilized single-molecule techniques measure equilibrium populations formed vitro by mixtures wild-type...
The huntingtin (HTT) protein transports various organelles, including vesicles containing neurotrophic factors, from embryonic development throughout life. To better understand how HTT mediates axonal transport and why this function is disrupted in Huntington's disease (HD), we study vesicle-associated find that it dimethylated at a highly conserved arginine residue (R118) by the methyltransferase 6 (PRMT6). Without R118 methylation, associates less with vesicles, anterograde trafficking...
Alpha-synuclein (alpha-syn) is a "natively unfolded" protein constituting the major component of intracellular inclusions in several neurodegenerative disorders. Here, we describe proteolysis experiments conducted on human alpha-syn presence SDS micelles. Our aim was to unravel molecular features micelle-bound using limited approach. The nonspecific proteases thermolysin and proteinase K, as well Glu-specific V8-protease, were used proteolytic probes. While at neutral pH easily degraded...
The aggregation of α-synuclein into amyloid fibrils constitutes a key step in the onset Parkinson's disease. Amyloid are major component Lewy bodies, histological hallmarks Little is known about mechanism α-synuclein. During this process, forms transient intermediates that considered to be toxic species. dimerization could represent rate-limiting protein. Here, we analyzed four covalent dimers α-synuclein, obtained by link N-terms, C-terms, tandem cloning two sequences and juxtaposition one...
Spinal and bulbar muscular atrophy is caused by polyglutamine (polyQ) expansions in androgen receptor (AR), generating gain-of-function toxicity that may involve phosphorylation. Using cellular animal models, we investigated what kinases phosphatases target polyQ-expanded AR, whether polyQ modify AR phosphorylation, how this contributes to neurodegeneration. Mass spectrometry showed preserve native phosphorylation increase at conserved sites controlling stability transactivation. In...